Hematopoietic potential correlates with Venus expression levels. (A) Hematopoietic progenitor numbers in E10.5 (30–36 SPs) CD31⁺ckit⁺V^med and CD31⁺ckit⁺V^high AGM sorted cells. Colony-forming unit-culture per embryo equivalent (CFU-C/ee) is shown, with colony types designated by colored bars. BFU-E, burst-forming unit erythroid; CFU-G/M/GM, CFU granulocyte, CFU macrophage, and CFU granulocyte-macrophage; CFU-GEMM, CFU granulocyte, erythroid, macrophage, megakaryocyte. The data represent the mean ± SEM of four independent experiments. (B) Percentage donor cell chimerism in recipient mice injected with CD31⁺ckit⁻V⁺, CD31⁺ckit⁺V⁻, CD31⁺ckit⁺V^med, or CD31⁺ckitV^high sorted E11 (41–49 SPs) AGM cells. Engraftment at 4 mo after transplantation was determined by flow cytometric analysis of Ly5.1/Ly5.2 marker expression of peripheral blood cells. Each dot represents one recipient receiving 1.3 to 4.1 embryo equivalent (ee) of sorted AGM cells. The data represent the mean ± SD. *, P ≤ 0.024; **, P = 0.0085; ***, P = 0.0003). (C) Overrepresentation of up-regulated differentially expressed genes (DEGs) in E10.5 CD31⁺ckit⁺V^med and CD31⁺ckitV^high sorted cells in IPA canonical pathways. (D and E) Mean FPKM values for genes in the Notch pathway (D) and heptad factor genes in E10.5 CD31⁺ckit⁺V^med and CD31⁺ckitV^high sorted E10.5 AGM cells (E). The data were compared using Student’s t test (*, P = 0.0404; **, P = 0.0096). The data represent the mean ± SEM of three independent experiments.